Journal: The Journal of Biological Chemistry

Article Title: Epithelial Mesenchymal Transition Induces Aberrant Glycosylation through Hexosamine Biosynthetic Pathway Activation *

doi: 10.1074/jbc.M116.729236

Figure Lengend Snippet: Comparison of surface glycan profile in A549 cells before and after induction of EMT by TGF-β. A, scheme representing binding specificities of the lectins used in this study. A549 cells were incubated without (control, Ct) or with 5 ng/ml of TGF-β for 48 h, and surface glycans were labeled with biotinylated lectins and incubated with FITC-conjugated avidin. B, histograms indicating the avidin (black line) binding, untreated control (blue lines) cells, and cells treated with TGF-β (red lines). C, bar graph comparing the fluorescence intensity for lectins in control and 48-h TGF-β-treated cells. The results are representative of three independent experiments. AAL, A. aurantia agglutinin; ECA, E. cristagalli agglutinin; PNA, peanut agglutinin; PSA, P. sativum agglutinin; SNA, S. nigra agglutinin; MAA, M. murensis agglutinin; L-PHA, phyto hemagglutinin L; E-PHA, phyto hemagglutinin E; VVL, V. villosa lectin; MIF, mean intensity of fluorescence. *, p < 0.01.

Article Snippet: The cell surface was blocked in a 3% BSA solution in PBS and incubated with the following biotinylated lectins: Sambucus nigra agglutinin, Erythrina cristagalli agglutinin, peanut agglutinin, Aleuria aurantia agglutinin, Pisum sativum agglutinin, Maackiaa murensis agglutinin, phyto hemagglutinin L, phyto hemagglutinin E, and Vicia villosa lectin (EY Laboratories) diluted in PBS-BSA in a concentration of 10 μg/ml.

Techniques: Comparison, Glycoproteomics, Binding Assay, Incubation, Control, Labeling, Avidin-Biotin Assay, Fluorescence